a new analytical method development and validation …

Rao et al. European Journal of Biomedical and Pharmaceutical Sciences

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A NEW ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF STABILITY

INDICATING RP-UPLC ASSAY FOR THE DETERMINATION OF LURASIDONE

HYDROCHLORIDE IN BULK AND TABLET DOSAGE FORMS.

Tirukkovalluri Siva Rao2*, Tene Sivaganesh

1,2, Kommula Ramasrinivas

1, Uppalapu Sesham Raju

1,

Pinninti Sateesh Kumar1

1Department of Analytical Chemistry, AurobindoPharma Ltd, Pydibhimavaram, India.

2Department of Inorganic & Analytical Chemistry, Andhra University, Visakhapatnam, 530003, India.

Article Received on 19/04/2018 Article Revised on 09/04/2018 Article Accepted on 30/05/2018

]

1. 0.INTRODUCTION

Lurasidone is a benzothiazol derivative (Figure 1)

belonging to second-generation antipsychotics class.It is

used for the treatment of schizophrenia and bipolar

depression. Although its mechanism of action is not fully

understood, it is believed that the efficacy of

LurasidoneHCl is mediated mainly through antagonist

activity at the dopamine D2, and the 5-

hydroxytryptamine (5HT, serotonin) receptors: 5-

HT2Aand 5-HT7.[1]

LurasidoneHcl demonstrated both

antipsychotic and antidepressant action. Despite its side

effects like higher rates of akathisia, Parkinsonism and

hyperprolactinemia, it has relatively lower risk for

developing sedation or overweight/ obesity.

LurasidoneHcl is available as tablets under the brand

name of Latuda. Several chromatographic methods have

been described for the quantification of Lurasidone Hcl

including spectrometry[2-5]

, TLC[6]

, HPTLC[7, 9]

,

GC/MS[8]

, HPLC[9-16]

and LC/MS.[17-22]

The goal of this

study was the development of a new UPLC method for

determination of LurasidoneHCl in its API bulk drug and

tablet dosage forms. Although there are some previous

HPLC methods developed for this purpose, they allow

analysis at a narrow concentration range and the

separation of stereoisomer’s not reported in any method.

Developed method has a wide concentration range, low

quantification and detection limit compare to all HPLC

and UV methods. It was validated according to the

international conference on harmonization guideline[23]

and applied for the analysis of bulk drug and tablet

dosage form.

SJIF Impact Factor 4.918 Research Article

ejbps, 2018, Volume 5, Issue 6 843-857.

European Journal of Biomedical AND Pharmaceutical sciences

http://www.ejbps.com

ISSN 2349-8870

Volume: 5

Issue: 6

843-857

Year: 2018

*Corresponding Author: Prof. Tirukkovalluri Siva Rao

Department of Inorganic & Analytical Chemistry, Andhra University, Visakhapatnam, 530003, India.

ABSTRACT

Method development, validation and stability indicating RP-UPLC method for Assay determination of

LurasidoneHcl in pharmaceutical products. This method has high degree of performance to separation and

quantification of Lurasidone even in the presence of its impurities. This efficient Separation can be achieved by

using new Waters Acquity CSH Phenyl hexyl with dimensions (100mm x 2.1 mm, 1.7μm). The buffer used in this

method KH2PO4 at pH 2.5 and Acetonitrile in the ratio of (65:35 v/v) with a flow rate of 0.5 ml/min and the

absorbance was monitored at 230 nm. The total run time was 3.0 min. The correlation coefficient of the method

shows good linear relationship with 0.9999. The limit of detection and quantification are determined for

LurasidoneHCl 0.01µg/ml and 0.03µg/ml. The signal to noise ratio has been observed 7 and 12 for LOD & LOQ

respectively. The precision of the method %RSD is less than 0.40% and the % recovery of LurasidoneHCl is

between 99.5 – 100.1 %.When the drug is subjected to different stress conditions and the resulting degradation

products obtained were not interfere during the determination of LurasidoneHCl.

KEYWORDS: Lurasidone HCl, RP-UPLC, Method development, Validation, Stability-indicating.


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(S) N

(R)

O

O

(R)(R)

N

N

SN

(Endo, 1R, 2S)-4-[2-(4-Benzo[d]isothiazol-3-yl-piperazin-1-ylmethyl)-cyclohexylmethyl]-4-aza-tricyclo[5.2.1.02,6]decane-3,5-dione

(S)

( R)

(S) N

(R)

O

O

(R)(R)

N

N

SN

(R)

(S)

LURASIDONE

(S) N

(R)

O

O

( S)( S)

N

N

SN

(R)

( S)

[(3aR, 4S, 7aS)(1S, 2S)-STEREO ISOMER

(S) N

(R)

O

O

(R)(S)

N

N

SN

(R)

(S)

[(3aR, 4S, 7aS)(1S, 2R)-STEREO ISOMER

(S) N

(R)

O

O

(R)(S)

N

N

SN

(R)

(S)

[(3aR, 4S, 7aS)(1R, 2S)-STEREO ISOMER

(S) N

(R)

O

O

(R)(R)

N

N

SN

( S)

(R)

[(3aS, 4S, 7aR)(1R, 2R)-STEREO ISOMER Fig: 1.

Table: 1.

Product Name Lurasidone Hydrochloride

Chemical Name

(3aR,4S,7R,7aS)-2[(1R,2R)-2-[4-(1,2-Benzisothiazol-3-Yl)- Piperazin-1-

Ylmethyl]Cyclohexyl Methyl]Hexahydro-4,7-Methano-2h-Isoindole-1,3-Dione,

Hydrochloride

CAS Reg.No: 367514-88-3

Molecular Formula C28H36N4O2S. HCl

Molecular Weight 529.14


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2.0. EXPERIMENTAL

2.1. Chemicals and reagents

Samples of Lurasidone HCl were contributed from

Aurobindo pharma Ltd, Pydibhimavaram, India. HPLC

grade Acetonitrile, AR grade KH2PO4 and

Orthophosphoric Acid were purchased from Merck india

Ltd, Mumbai. High pure water was prepared by using

Millipore Milli-Q water purification system. New Waters

Acquity CSH Phenyl hexyl (100 x 2.1 mm, 1.7 μm)

column (Part no. # 186005407), waters AcquityBEH

(50mm×2.1mm, 1.7μm), waters Acquity HSS

T3(100mm×2.1mm,1.8μm) and waters Acquity HSS

C18(100mm×2.1mm,1.8μm) was procured from Waters

India Ltd, Bangalore.

2.2. Instrumentation (Apparatus)

Acquity UPLC system equipped with an LC pump

(model ACQ-BSM) used for method development and

validation itscontain, an online degasser, auto sampler

(model ACQ-SM) with thermostat, and detector (TUV)

(model ACQ-TUV). The data was acquired, monitored

and processed by using Empower3 software. Design

expert version 9 (Stat-Ease Inc., Minneapolis, USA) was

used for the optimizing chromatographic conditions. The

buffers pH was monitored by using Metrohm 780 p

H

meter and weights taken by using the Sartorius

CPA225D balance. In this research work we have

attempted various chromatographic columns for method

development. After number of trails we have been

selected Waters Acquity CSH Phenyl hexyl (100 x 2.1

mm, 1.7 μm) column based on its performance.

2.3. Chromatographic conditions

The chromatographic separations were performed by

using New Waters Acquity CSH Phenyl hexyl (100 x 2.1

mm, 1.7 μm) (Part no. # 186005407). The pH

of the

buffer KH2PO4was adjusted to pH

2.5 by addition of

orthophosphoric Acid whereas the mobile phase contains

buffer (pH 2.5) and Acetonitrile in the ratio of (65:35).

The flow rate of the mobile phase is 0.5 mL/min. The

column temperature was maintained at 50°C and the

absorption was measured at 230nm. The total Run time

of the method was found to be 3.0min and the injection

volume was 6µL. Water and Acetonitrile in the ratio of

(50:50 v/v) used as a diluent. Mobile phase and diluent

Filter through 0.22 finer porosity membrane filter. The

chromatographic conditions were given in table-2.

Table: 2

Instrument : RP-UPLC make by Waters

Mode of analysis : Isocratic

Flow rate : 0.5 mL/min

Detector wave length : 230 nm

Column temperature : 500C

Injection volume : 6.0 µL

Column : WatersAcquityCSHPhenylhexyl(100x2.1mm, 1.7μ)

Run time : 3.0 min

Sample Manager Temp : Ambient

Inference: Results from the table-2 indicated that the

eluting run time was completed within 3 minutes

compared with other HPLC methods for assay

determination in the literature. Moreover the injection

volume of the sample required for RP-UPLC method is 6

µL is sufficient but in the case of HPLC methods it needs

about 20 µL.

Table: 3

S. No. NAME RT

1 Lurasidone HCl ~2.0

2.4. Preparation of standard and sample solutions

Preparation of standard solution

Weighed and transferred accurately 50 mg of

LurasidoneHCl working standard into a 100 mL clean,

dry volumetric flask add 50 mL of diluent and sonicate

to dissolve. Make up to volume with diluent. Diluted 5

mL of this solution to 50 mL with diluent. Filter through

0.22 finer porosity membrane filter.

Preparation of sample solution

Weighed and transferred accurately 50 mg of

LurasidoneHCl sample into a 100 mL clean, dry

volumetric flask add 50 mL of diluent and sonicate to

dissolve. Make up to volume with diluent. Diluted 5 mL

of this solution to 50 mL with diluent. Filter through

0.22 finer porosity membrane filter.

2.5. Calculation of Lurasidone HCl Assay

The samples assay was calculated by the following

equation

Where AT is peak average area due to LurasidoneHCl in

the sample preparation, AS is Average peak area due to

Lurasidone HCl in the standard. WS is the weight of

Lurasidone HCl standard taken in mg, WT is the Weight

of the sample taken in mg, P is the Potency of the

Lurasidone HCl working/reference standard. W.C is the

water content of the Lurasidone HCl.

).100(

100

5

50100

50

5

100 CWP

WT

WS

AS

AT


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3.0. RESULTS AND DISCUSSION

3.1.0. Method development and Optimization of RP-

UPLC conditions

The RP-UPLC conditions were optimized by using trials

with different columns, several mobile phase

compositions; flow rate and pH

were studied. The

Analytical method development and validation play an

important role in the determination of Assay in

pharmaceutical products. In this method less quantity of

solvents are used and total consumption of solvents is not

more than 0.525ml per run time. One of the principles of

green chemistry is prevention of waste. Further

advantages of RP-UPLC method is able to increase the

speed, sensitivity and resolution compared to HPLC

methods. By considering above aspects, RP-UPLC

instrument is a most suitable technique for the assay

determination of Lurasidone HCl.

3.1.1. Selection of stationary phase

Based on the structure, molecular weight of API and

impurities present in the products C18 columns like

Waters BEH C18 column were initially screened for the

separation. But these columns failed to provide

acceptable separation and peak shape. For also several

other HSS C18 columns with other stationary phases are

screened for separation but a remarkable selectivity was

achieved with New Waters Acquity CSH Phenyl hexyl

(100mm x 2.1 mm, 1.7 μm) partial size was finalised.

This column was not yet used in any other methods.

Waters AcquityBEH C18 column; The trifunctionally

bonded BEH Column particle gives a widest usable pH

range i.e’s 1-12range, superior low pH stability and ultra

low column bleed for high sensitivity applications.

Waters Acquity CSH Phenyl hexyl column; this

column are used to provide an alternative selectivity and

are a valuble tool for method development. The

trifunctionally bonded C6phenyl ligond is a robust and

low bleed sorbents that selectively retains poly aromatic

compounds through Π-Π interactions.

Table: 4 Stationary phases Information.

Brand Acquity UPLC Particle size(dp) 1.7μm

%Carbon Load 14 Particle Substrate Hybrid

Bonding Technology Phenyl-Hexyl Pore size 130Å

Chemistry Phenyl Silanol Activity Low

Endcapped yes Surface Area 185

ID 2.1 mm Technology CSH

Length 100mm USP Classification L11

Particle Shape Spherical Units in package 1/pkg

Mode Reversed-phase pHrange 1-11

3.1.2. Selection of Mobile phase

As one of the objectives of the method is to develop a

different buffer, i.e’s Ammonium acetate buffer with pH

9.0, diammonium hydrogen orthophosphate with pH3.3

and potassium dihydrogen orthophosphate+water buffers

were evaluated. It is observed that in potassium

dihydrogen ortho phosphate buffer is a promising

candidate for efficient separation of impurities. The

organic modifiers, methanol and acetonitrile were used at

different composition conditions. Based on the results,

acetonitrile was finalised as a organic modifier.

3.1.3. Selection of Diluent

LurasidoneHCl was practically insoluble in water and

slightly soluble in methanol. But It is soluble in

acetonitrile+ water composition and stable for at least 24

hrs at 25°C. Hence, acetonitrile + Water (50:50 v/v) was

selected as a diluent.

3.1.4. Experimental design for optimising flow rate,

buffer concentration and column temperature

In initial method development trials with one factor at a

time (OFAT) variation revealed that the flow rate and

column temperature and composition of organic modifier

had significant impact on selectivity. Since optimising

the chromatographic parameters with OFAT approach

consumes lot of time and does not provide the design

space, a design of experiments (DoE) was used for

optimising these chromatographic parameters. The

design space defines the experimental region in which

changes to method parameters will not significantly

affect the quality and results. As working within the

design space is not considered as a change, the scientist

can have freedom to operate the method at different

chromatographic condition.

Based on the analysis, it was understood that, to obtain

good analyte peak shape, column temperature and flow

rate are monitored at 50°C and 0.5 mL/min respectively.


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Table: 5 Optimized chromatographic conditions.

Instrument : RP-UPLC make by Waters

Mode of analysis : Isocratic

Flow rate : 0.5 mL/min

Detector wave length : 230 nm

Column temperature : 500C

Injection volume : 6.0 µL

Column : WatersAcquityCSHPhenylhexyl (100x2.1mm, 1.7μ)

Run time : 3.0 min

Sample Manager Temp : Ambient

3.2.0. METHOD VALIDATION

The developed analytical method was subjected to

validation with respect to various parameters such as

linearity,limit of quantification (LOQ), limit of detection

(LOD), accuracy, precision, recovery studies, specificity

and reproducibility as per the ICH guidelines.

3.2.1. SYSTEM SUITABILITY

In the optimized RP-UPLC conditions, system suitability

parameters were evaluated for Lurasidone

HCl(Fig2).Tailing factor for Lurasidone HCl was not

more than 2.0. The USP plate count for Lurasidone HCl

is not less than 2000. % RSD of the six injections was

not more than 1.0%.The resolution between

LurasidoneHCland sterioisomer-1 is not less than 2.0 and

also resolution between LurasidoneHCland sterioisomer-

2 is not less than2.0.The resultsare summarised in

Table: 6.

System suitability chromatogram

Fig: 2.

Table: 6

Parameters LurasidoneHCl

Limit Initial SST Bracketing SST

Tailing factor 1.28 1.28 NMT 2.0

USP Plate count 4430 4464 NLT 2000

Retention time (min) 1.99 1.99 ~2.0

Resolution between LurasidoneHCland

sterioisomer-1 2.70 2.58 NLT 2.0



3.2.2. SPECIFICITY

The specificity of the proposed method was

demonstrated by interference study. It was found that

presence of some common exicipients did not

interferences at the retention time of LurasidoneHCl.

Thus the developed method can be successfully applied

for determination of LurasidoneHCl in bulk drug and

tablet dosage form.

3.2.3. Blank and Impurity interference

The blank, sample enriched with impurities were

prepared and injected in RP-UPLC. No interference was

observed at any of the peaks of interested in blank.


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Blank chromatogram

Fig:3

3.2.4. Forced degradation/ Stress study

In order to determine whether the analytical method and

assay were stability-indicating, LurasidoneHCl bulk drug

and tablet dosage form was stressed under various

conditions to conduct forced degradation studies.

Lurasidone HCl was soluble in acetonitrile+water in the

composition of (50:50). All solutions for forced

degradation studies were prepared to yield concentration

50μg/g of LurasidoneHCl.

Acid degradation (5N HCl)

Solution of Lurasidone HCl (50μg/g) for acid

degradation study was prepared using 5N hydrochloric

acid solution and the resultant solution was 1 hours to

facilitate acid degradation of Lurasidone HCl.

Alkali degradation (5N NaOH)

Solution of Lurasidone HCl (50μg/g) for base

degradation study was prepared by using 5N NaOH

resultant solution was 1 hours to facilitate alkali

degradation of Lurasidone HCl.

Oxidation (30 % H2O2)

Solution of Lurasidone HCl (50μg/g) for peroxide

degradation study was prepared using 30% H2O2

resultant solution was 1 hour to facilitate peroxide

degradation of Lurasidone HCl.

Thermal degradation

LurasidoneHCl bulk drug was kept in hot oven for

24hours at 80°C to determine the effects of thermal

degradation on the stability of Lurasidone HCl by using

the analytical method.

Photo stability (Sun light)

Lurasidone HCl bulk drug was exposed to sunlight to

determine the effects of light irradiation on the stability

of LurasidoneHCl in the solid state. Approximately 200

mg of Lurasidone HCl bulk drug was spread on a glass

dish in a layer that was less than 2 mm thick. All samples

for photo stability testing were placed indirect sunlight

exposed for 48hours. The bulk drug were removed from

the sun light and the contents of bulk drug equivalent to

100 mg of LurasidoneHCl was accurately weighed and

transferred to volumetricflask to make the final

concentration 50μg/g of Lurasidone HCl. The proposed

method evaluates the ability to separate the impurities of

Lurasidone HCl from its degradation products shown in

Figure: 4 to 9. The results and system suitability

parameter are summarised in Table: 7

Undegraded Sample chromatogram. Sample chromatogram of peroxide degradation

Fig:4 Fig:5


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Sample chromatogram of Thermal degradation Sample chromatogram of Photo degradation

Fig:6 Fig:7

Sample chromatogram of Acid degradation Sample chromatogram of Base degradation

Fig:8 Fig:9

Table:7

Parameters LurasidoneHCl Limit

Initial SST Bracketing SST








Degradation study

S.No Condition Time % Degradation

% Assay %Degradation 1 Undegraded sample Fresh 99.9% NA 2 5N HCL added sample 60min 35.3% 64.7% 3 5N NaOH added sample 60min 38.1% 61.9% 4 30% H2O2 added sample 60min 49.8% 50.2% 5 Thermal 80°C heated sample 24 hrs 99.6 0.4% 6 Photo Degradation sample 48 hrs 99.3 0.7%

Inference: Based on the above results from the table it is

revealed that in the HCl, Oxidative and NaOH

degradation process high degradation products are

obtained than that of the all undegarded samples.

3.2.5. LINEARITY

3.2.6. Linearity for LurasidoneHCl Assay

The calibration curve was plotted over the concentration

range of 0.03 to 75.3μg/g of Lurasidone HCl. The

calibration curves were prepared by plotting the peak

area versus the concentration and analyzed through linear

regression (Figure-10). The linearity was observed in the


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expected concentration range, demonstrating its

suitability for analysis.The system suitability and results

are summarised in Table: 8.

Table: 8 Linearity (Correlation coefficient)










Precision

Injection-01 1208217

%RSD NMT 1.0%






Mean 1206957

SD 1178

RSD 0.10

S.No Concentration(μg/ml) Response(Area)

1 75.33 1761924

2 60.3 1436463

3 50.22 1207821

4 40.2 958124

5 25.11 598262

6 15.1 350948

7 5.022 115780

8 2.511 66112

9 1.0044 33180

10 0.5022 11801

11 0.2511 10328

12 0.062775 2714

13 0.030388 1374

14 0.010506 844

Slope 23611.2340199

Intercept 3773.6523776

Correlation 0.99988

LurasidoneHCl Linearity Calculation sheet

Fig:10


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Inference: Based on correlation coefficient values from

table-8 were within the Acceptance criteria.

3.2.7. LIMIT OF DETECTION AND LIMIT

QUANTIFICATION

The limit of detection (LOD) and limit of quantitation

(LOQ) were established for LurasidoneHClby diluting

the standard stock solution. The concentration at

0.01µg/g, 0.030µg/g LOD and LOQ for LurasidoneHCl.

The signal to noise ratios was found to more than 7 and

12 respectively for the analyte. Hence these

concentrations were finalised as LOD and LOQ

concentrations. Further precision was found to be 2.67%

at LOD level for analyte and 4.06 at LOQ level the

results are summarised in Table: 9. Overlay

chromatograms of LOD & LOQ are representing below.

Table: 9 LOD and LOQ Precision.






Resolution between

LurasidoneHCland sterioisomer-1 2.56 2.57 NLT 2.0

Resolution between

LurasidoneHCland sterioisomer-2 2.55 2.50 NLT 2.0

Precision


%RSD NMT 1.0%






Mean 1205744

SD 1914

RSD 0.16

LurasidoneHCl LOQ (%RSD NMT 10.0%) LOD (%RSD NMT 33.0%)

Injection-1 1280 710






Average 1240 742

Standard Deviation 50 20

%RSD 4.06 2.67

Overlay Chromatogram of LOD

Fig-11


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Overlay Chromatogram of LOQ

Fig-12

3.2.8. ACCURACY

3.2.9. Accuracy for Assay of LurasidoneHCl

The accuracy of the method was determined by

calculating recoveries of Lurasidone HCl by the standard

addition method. Known amounts of standard solutions

of Lurasidone HCl (80, 100, and 120 % level) were

added to previously analyzed sample solutions of bulk

drug. The percentage of recoveries was calculated. The

percentages of recoveries were between 98.0 to102.0.The

results are summarised in the following Table: 10.

Table: 10 Accuracy and SST parameters.










Precision


RSD (NMT 1.0%)






Mean 1216385

SD 1628

%RSD 0.13

% of Drug Added Amount Added

(W/W)

Amount Recovered

(W/W)

% Recovery

(Criteria 98.0 to 102.0)

LurasidoneHClat 80% 80.3 80.0 99.7










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Inference: Results from the table-10, it is illustrated that

recovered concentration of spiked samples was found to

be within the acceptance criteria i.e. 98.0 to 102.0.

3.2.10. PRECISION

The precision of the method was demonstrated by system

precision and method precision.

3.2.11. System precision

System precision for assay was demonstrated by

injecting standard solution under the same operating

conditions. The peak areas of Lurasidone HCl were

measured and the % RSD was found to be 0.19%.The

results are summarised in Table: 11.

Table: 11

Parameters Lurasidone HCl





Resolution between Lurasidone HCl and


Resolution between Lurasidone HCl and


System precision

S.No Area Criteria % RSD

Injection-1 1225499

(NMT1.0%)

Injection-2 1224730

Injection-3 1225963

Injection-4 1223581

Injection-5 1224343

Injection-6 1223344

Mean 1224577

SD 10.36

%RSD 0.08

Inference: Results from the table-11, it is illustrated

%RSD of standard was found to be within the acceptance

criteria i.e. NMT 1.0%

3.2.12. Method precision

Method precision for analyte was demonstrated by

preparing six samples at spec level. These solutions were

injected along with a standard solution of Lurasidone

HCl prepared at spec level. The relative standard

deviation all six preparations results was found to be

0.23%.The results are summarised in Table: 12.

Table: 12

Method precision

Lurasidone HCl Assay result % RSD Criteria % RSD

Sample-01 99.6

0.23% NMT 1.0%

Sample-02 99.3

Sample-03 99.5

Sample-04 99.8

Sample-05 99.2

Sample-06 99.3

Inference: Results from the table-12, it is illustrated that

%RSD of results precision was found to be within the

acceptance criteria i.e. NMT 1.0%

3.2.13. Intermediate precision (ruggedness):

Intermediate precision for analyte was demonstrated by

preparing six different samples at spec level by different

analyst and different day. These solutions were injected


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along with a standard solution of Lurasidone HCl

prepared at spec level. The relative standard deviation of

analyte content obtained from all six preparations results

was found to be 0.41%. The system suitability

parameters and results are summarised in Table: 13.

Table: 13










System Precision


%RSD (NMT 1.0%)






Mean 1212006

SD 1667

%RSD 0.14

Intermediate precision


Sample-01 100.2

0.33% NMT 1.0%

Sample-02 99.7

Sample-03 100.6

Sample-04 99.8

Sample-05 99.9

Sample-06 99.9

Cumulative RSD for precision

Analyst-01 Analyst-02 Cumulative (12 Results) Criteria % RSD

0.23% 0.33% 0.40% NMT 1.0

Inference: Sample Results from the table-12, 13 it is

observed that cumulative RSD’s were found to be within

the limit i.e. NMT1.0%.

3.3. FILTER COMPATIBILITY

Filter compatibility to the sample is concluded from the

recovery study indicated that there is no absorption of

these component to filter.

3.4. SOLUTION STABILITY

The standard and samples solutions were kept on room

temperature at 25°c and injected the aged samples (Every

one hour) into the RP-UPLC. The peak areas

corresponding to Lurasidone HCl were measured.

Calculated the similarity factor and found that the values

are below 1.0% RSD. Thus indicates the sample and

standard solutions are stable for at least 24 hrs when

stored on room temperature condition. System suitability

parameters are shown below table-14.

Table-14






Resolution between Lurasidone

HCl and sterioisomer-1 2.65 2.50 NLT 2.0

Resolution between Lurasidone

HCl and sterioisomer-2 2.57 2.49 NLT 2.0


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3.5. METHOD ROBUSTNESS

Method robustness was performed by applying small

changes in the ratio of mobile phase, injection volume,

and column temperature, pH and flow rate. The results of

change in ratio of mobile phase, column temperature,

wavelength, and injection volume are shown in Table 15.

The flow rate 0.45– 0.55 mL/min, buffer pH 2.40 to

2.60,wavelength 227 to 233nm and Column temperature

45°C to 55°C and Study of the mobile phase composition

61.5:38.5, 68.5:31.5. Based on this, the method is proved

to be robust and can easily be implemented in quality

control laboratories for the regular analysis of

Lurasidone HCl samples with great confidence.

Table-15.

Method Robustness

Ch

rom

ato

gra

ph

ic P

ara

met

er

Co

nd

itio

n

Ret

enti

on

tim

e(R

T)

~2

.0

Resolution

Th

eore

tica

l p

late

s(N

) N

LT

20

00

Ta

ilin

g(T

) N

MT

2.0

Res

ult

(NL

T 9

8.0

TO

10

2.0

)

Lurasidone

HCl&Sterioiso

mer-1

Lurasidone

HCl&Steriois

omer-2

Init

ial

Aft

er

Init

ial

Aft

er

Wavelength(nm) 227 1.97 2.66 2.62 2.64 2.59 4561 1.33 100.0

233 1.97 2.65 2.60 2.62 2.62 4492 1.32 100.2

Temperature(°C) 45°C 1.97 2.59 2.61 2.67 2.67 4397 1.34 99.4

55°C 1.96 2.61 2.62 2.60 2.60 4558 1.31 99.8

Acetonitrile (%) 61.5:38.5 1.41 2.12 2.10 2.07 2.04 4341 1.41 99.7

68.5:31.5 2.5 3.19 2.84 3.12 3.15 4993 1.30 99.9

Flow rate(ml/min) 0.45 2.18 2.67 2.65 2.63 2.63 4519 1.35 100.0

0.55 1.81 2.61 2.63 2.59 2.60 4395 1.29 100.6

pH variation 2.40 1.96 2.69 2.56 2.71 2.60 4433 1.32 100.0

2.60 1.97 2.65 2.66 2.66 2.62 4458 1.32 99.9

Inference: Solvent composition is more critical based on

above data. Measure the mobile phase composition

accurately.

4.0. BATCH ANALYSIS

20 tablets of Lurasidone HCl with brand name of Sun

pharmaceuticals tablets was purchased and analyzed with

equal quantity of 3samples. Each tablet having around

135mg it contains about 40mg of Lurasidone HCl API.

20 tablets were taken and crush the samples with the help

of mortar and analysed with optimum chromatographic

conditions. But the sample solution slight haziness

observed it’s filtered with 0.22µ filter paper and

analysed. The SST parameters and results are

summarized in table-16.

DETAILS OF TABLET

Marketed By

Sun Pharma laboratories limited,

Andheri (E), Mumbai-400059,

B.No:DT1610036B,

Unit: II.

Table-16.

Parameters Lurasidone HCl





Resolution between Lurasidone HCl and sterioisomer-1 2.54 2.58 NLT 2.0

Resolution between Lurasidone HCl and sterioisomer-2 2.57 2.52 NLT 2.0

TABLETS ANALYSIS RESULTS


Tablet-01 100.0

0.28% NMT 1.0% Tablet-02 100.2

Tablet-03 100.6


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5.0. DISCUSSION

The purpose of the present work was to develop a short,

robust, precise RP-UPLC method for the accurate

quantisation of Lurasidone HCl Assay. This method was

developed for drug substance and drug product also. As

mentioned in the introduction section, several reports are

available for quantification of Lurasidone HCl but

present method was highly sensitive.The developed

method was successfully validated for drug substance

and drug product as per the ICH guideline. The proposed

method is much superior to reported methods in terms of

solvent consumption, run time, instrumental technique

(RP-UPLC), selectivity, and applicability to Assay

analysis.

6.0. APPLICATION TO PHARMACEUTICAL

INDUSTRY

This work will help industry to develop, manufacture and

launch the product in a fast and economical way which

in turn reduces the cost of the medicine and help the

patient to avail quality, innovative and affordable

medicine.

7.0. CONCLUSION

A stability indicating RP-UPLC Assay method has been

developed for determination of Lurasidone HCl.

Developed method was proved to be robust using the

experimental design, this method can be successfully

implemented in the quality control lab for the routine

analysis of this product. Further this RP-UPLC method

was successfully validated as per ICHQ2 (R1) guideline

and proved to be precise, linear, sensitive, accurate, and

robust. This method is short and simple, hence

implementation of this method in quality control and

analytical development labs can yield high throughput.

As low amounts of solvents are required, implementation

of this method will be eco friendly. This was the first

RP-UPLC Assay method that can accurately quantitative

the Lurasidone HCl Assay in drug substance and drug

product.

ACKNOWLEDGEMENTS

The authors are heartily thankful to acknowledge

Aurobindo Pharma Ltd and Andhra University for

providing all the facilities to carry out the research work.

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